Characterisation of GLUT4 trafficking in HeLa cells: comparable kinetics and orthologous trafficking mechanisms to 3T3-L1 adipocytes

PeerJ

article

Silke Morris¹  Niall D. Geoghegan²  Jessica B.A. Sadler¹  Anna M. Koester¹  Hannah L. Black³  Marco Laub¹  Lucy Miller¹  Linda Heffernan⁴  Jeremy C. Simpson⁴  Cynthia C. Mastick⁵  Jon Cooper²  Nikolaj Gadegaard²  Nia J. Bryant³  Gwyn W. Gould⁶ 

[1] Institute of Molecular Cell and Systems Biology, University of Glasgow;School of Engineering, University of Glasgow;Department of Biology, University of York;School of Biology & Environmental Science, University College Dublin;Molecular Biosciences, University of Nevada - Reno;Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde

关键词: Membrane;    Transport;    Insulin;    Diabetes;    GLUT4;    Endosome;   

DOI  :  10.7717/peerj.8751

学科分类：社会科学、人文和艺术（综合）

来源: Inra

【 摘 要 】

Insulin-stimulated glucose transport is a characteristic property of adipocytes and muscle cells and involves the regulated delivery of glucose transporter (GLUT4)-containing vesicles from intracellular stores to the cell surface. Fusion of these vesicles results in increased numbers of GLUT4 molecules at the cell surface. In an attempt to overcome some of the limitations associated with both primary and cultured adipocytes, we expressed an epitope- and GFP-tagged version of GLUT4 (HA–GLUT4–GFP) in HeLa cells. Here we report the characterisation of this system compared to 3T3-L1 adipocytes. We show that insulin promotes translocation of HA–GLUT4–GFP to the surface of both cell types with similar kinetics using orthologous trafficking machinery. While the magnitude of the insulin-stimulated translocation of GLUT4 is smaller than mouse 3T3-L1 adipocytes, HeLa cells offer a useful, experimentally tractable, human model system. Here, we exemplify their utility through a small-scale siRNA screen to identify GOSR1 and YKT6 as potential novel regulators of GLUT4 trafficking in human cells.

【 授权许可】

CC BY